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Fungi Associated with Pistacia Integerrima with a Description of a New Species and One New Record from India

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Fungi Associated with Pistacia Integerrima with a Description of a New Species and One New Record from India Acta Mycologica DOI: 10.5586/am.1100 ORIGINAL RESEARCH PAPER Publication history Received: 2017-04-01 Accepted: 2017-07-03 Fungi associated with Pistacia integerrima Published: 2017-12-29 with a description of a new species and one Handling editor Tomasz Leski, Institute of Dendrology, Polish Academy of new record from India Sciences, Poland Authors’ contributions Ajay Kumar Gautam1*, Shubhi Avasthi2 AKG: research idea, conducting 1 experiments, manuscript School of Agriculture, Faculty of Science, Abhilashi University, Mandi 175028, Himachal Pradesh, preparation; SA: manuscript India 2 preparation, reviewing drafts of Department of Botany, Abhilashi Post Graduate Institute of Sciences, Ner Chowk, Mandi 175008, the paper Himachal Pradesh, India * Corresponding author. Email: [email protected] Funding The research has been conducted on authors own expenses. Abstract Pistacia integerrima is a deciduous tree species belonging to the family Anacardiaceae. Competing interests No competing interests have Te plant possesses numerous phytochemicals of ethno-medicinal importance. In been declared. a routine mycological survey carried out from July 2013 to June 2014, leaves of P. integerrima were found infected with fungi causing rust and blight diseases. Te Copyright notice morphological and microscopic observations revealed three fungi, namely Skierka © The Author(s) 2017. This is an himalayensis, Pestalotiopsis sp., and Pileolaria pistaciae, which were found to cause Open Access article distributed under the terms of the Creative rust and blight diseases. One new species of rust fungi, namely Skierka himalayensis Commons Attribution License, sp. nov., and Pestalotiopsis sp. are reported for the frst time from India. Te detailed which permits redistribution, descriptions and illustrations of these three phytopathogenic fungi are provided in commercial and non- this paper. commercial, provided that the article is properly cited. Keywords Citation India; leaf blight; new report; Pistacia integerrima; rust; taxonomy Gautam AK, Avasthi S. Fungi associated with Pistacia integerrima with a description of a new species and one new record from India. Acta Mycol. Introduction 2017;52(2):1100. https://doi. org/10.5586/am.1100 Pistacia integerrima J. L. Stewart ex Brandis is a tree species native to Asia. Te plant belongs to family Anacardiaceae. Te common names for this species in Hindi are Digital signature kakar singhi, kakra, and kakring. It is a large deciduous tree having numerous ethno- This PDF has been certifed using digital signature with a trusted timestamp to medicinal properties. Traditionally, the plant parts are used to treat respiratory distress, assure its origin and integrity. A verifcation trust dialog appears on the PDF document all types of coughs, jaundice, chronic wounds, dysentery, and also have the antiseptic when it is opened in a compatible PDF properties [1–5]. reader. Certifcate properties provide further details such as certifcation time Pistacia integerrima is reported to sufer from a number of fungal diseases. Dur- and a signing reason in case any alterations made to the fnal content. If the certifcate ing a routine mycological survey In India carried out from July 2013 to June 2014, is missing or invalid it is recommended to two rust and a blight disease on P. integerrima caused by diferent fungal pathogens verify the article on the journal website. were noticed. All rust and blight samples were examined in detail for morphological characteristics. In the course of identifcation of these fungi, two were reported as new records from India. Material and methods Specimen sampling Te specimens infecting leaves of Pistacia integerrima were collected in diferent seasons, specifcally: rust samples during the prerainy (May–July) and winter (October–De- cember) season, while leaf blight samples in postrainy (August–October) season. Te collection site – Mandi, is located in Himachal Pradesh. Te area is a well-known region Published by Polish Botanical Society 1 of 6 Gautam and Avasthi / Fungi associated with Pistacia integerrima of the state and popular for its fertile land as well as plain and hilly regions. Specimens collected were studied in a laboratory and deposited at School of Agriculture, Faculty of Science, Abhilashi University, Mandi, Himachal Pradesh, India. Fungal isolation and identifcation Standard methods for collection, preservation, and description of fungi have been fol- lowed. For examination of morphological features of rusts, the specimens were mounted in lactophenol and examined with an Olympus BH-2 microscope. For generic assign- ment of rust species, the manual of Cummins and Hiratsuka [6] was consulted. For examination of morphological features of Pestalotiopsis, the material was cultured. For this purpose, twigs and young leaves showing blight symptoms were randomly collected, put in polythene bags, and transported to the laboratory for further investiga- tion. Small dissected infected leaf tissues were surface sterilized in 95% ethanol (20 s) followed by 0.525% NaOCl (60 s) and then washed three times with sterilized distilled water. Te leaf pieces were incubated aseptically on potato dextrose agar (PDA; Hi Media, India). Te cultures were maintained at 25 ±2°C for 12 hours. Fungi that grew from lesions were subcultured on PDA containing 50 mg/L streptomycin sulfate to maintain single species cultures. Subsequently, microscopic and cultural examinations were carried out for pathogen identifcation. Results – taxonomy Skierka himalayensis A. K. Gautam & S. Avasthi, sp. nov. (Fig. 1a,b) Mycobank: MB 821919. Description. Leaf spots hypophyllous, initiated as small brown to blackish, rounded rust sori, surrounded by a reddish yellow or chlorotic zone. Te pustules initially scat- tered, later coalesced to form a hard dry crust on the leaf surface. Telia subepidermal; teliospores one-celled, sessile, 35.7–48.3 (mean ±SD, 41.26 ±3.99) × 10.5–18.9 (mean ±SD, 14.28 ±3.1) μm in size, walls 2.5–3.5 μm thick; teliospores produced in irregular succession, strongly adherent and extruded in long, hair-like columns, germ pore not observed. Fig. 1 Skierka himalayensis. a Leaf showing rust infection. b Teliospores seen in LM. Scale bar: 10 μm. © The Author(s) 2017 Published by Polish Botanical Society Acta Mycol 52(2):1100 2 of 6 Gautam and Avasthi / Fungi associated with Pistacia integerrima Type. India, Himachal Pradesh: Mandi, elev. ca. 760 m a.s.l. (2495 f), on leaves of Pistacia integerrima (Anacardiaceae), November 24, 2014, leg. A. K. Gautam (holotype: AUMH 1097). Host and distribution. On Pistacia integerrima (Anacardiaceae), India. The rust symptoms were noticed on mature leaves during May 2014. Te disease symptoms were observed only on mature leaves. Notes. Tere are 13 Skierka species reported on diferent hosts across the world (Index Fungorum, http://www.indexfungorum.org, accessed on April 1, 2017). Te species newly described here, Skierka himalayensis, is easily distinguishable from morpho- logically similar species, namely S. canarii Racib. [7] and S. petchii (Syd.) Mains [8]. Skierka canarii has smaller teliospores (64–110 µm long) with thinner wall (1.5–2.5 µm). Similarly, broader length range of teliospores, greater wall thickness at sides and apex, and shorter beak size has been observed in S. petchii (Syd.). Te morphological characters of all reported species of Skierka are presented in Tab. 1. Tab. 1 Te morphological characters of Skierka species. Species Teliospores References Skierka himalayensis A. K. Gautam & 10.5–18.9 (mean ±SD, 14.28 ±3.1) × 35.7–48.3 (mean ±SD, 41.26 Present study S. Avasthi ±3.99) μm; wall 2.5–3.5 μm thick Skierka congensis Henn. 7–10 × 60–80 µm [14] Skierka philippinensis Mains 11–19 × 64–110 µm; wall smooth, hyaline, 1.5–2.5 µm thick [15] Skierka canarii Racib. 11–19 × 64–110 µm; wall smooth, hyaline, 1.5–2.5 µm thick [7] Skierka diploglottidis (Cooke & Mas- 15–18 × 70–90 µm; wall colorless, 1.5 µm thick [15] see) Mains Skierka petchii (Syd.) Mains 12–18 × 38–44 µm, the apex long attenuate, 25–50 µm long; wall hya- [8,15] line, inner wall 1 µm thick Skierka agallocha Racib. 8–12 × 60–100 µm; wall smooth, thin-walled, the apex 18–25 µm [16] Skierka robusta Doidge 20–27 × 120–180 µm; acuminate, elongated into a long fliform pro- [17] cess; wall 3.0–3.5(–5.0) µm thick Pestalotiopsis sp. Steyaert, Bull. Jard. bot. État Brux. 19: 300 (1949) (Fig. 2a–c, Fig. 3a–c) Description. Mycelial growth on potato dextrose agar (PDA) cottony white; conidia produced in ink-like fruiting bodies. Hyphal tips of the fungus were inoculated on PDA medium, where acervular conidiomata (22.8–29.1 × 5.3–8.9 μm) were developed afer 2 weeks of inoculation. Conidia fve-celled, smooth, with hyaline cylindrical to conical apical and basal cells; middle conidial cells dark brown with two upper cells sometimes darker; conidial size variable, 18.7–28.6 (mean ±SD, 24.82 ±3.26) × 5.3–9 (mean ±SD, 6.05 ±1.1) μm; both apical and basal cells bear appendages: three apical appendages, 6.6–17.6 (mean ±SD, 9.35 ±3.52) μm long and single basal appendage, 3.85–7.7 (mean ±SD, 5.33 ±1.07) μm long. Based on morphological characters described above, the fungus was identifed as Pestalotiopsis sp. [9]. Material investigated. India, Himachal Pradesh, Mandi, ca. 760 m a.s.l. (2495 f), on leaves of Pistacia integerrima (Anacardiaceae), September 9, 2013, leg. A. K. Gautam (AUMH 1015). Hosts and distribution. On Pistacia integerrima, Corylus avellana L. (Betulaceae), Juglans regia L. (Juglandaceae); India, Turkey. In India, the frst symptoms of the © The Author(s) 2017 Published by Polish Botanical Society Acta Mycol 52(2):1100 3 of 6 Gautam and Avasthi / Fungi associated with Pistacia integerrima Fig. 2 Pestalotiopsis sp. a,b Leaves showing blight symptoms. c Mycelial colony grown on potato dextrose agar for 10 days. disease on Pistacia integerrima were noticed during August–September 2013. Interestingly, the disease symptoms were observed only on young twigs or leaves.
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